Fig.1. Cold/β3-agonism induces a fibrogenic-to-adipogenic phenotypic shift in adipose stromal cells.

(A-E) Young (2-month-old) and aged (12-month-old) mice were acclimated to thermoneutrality and treated with vehicle or CL for 4 days to induce beiging.

(A) Immunofluorescence staining for UCP1 (green) in iWAT. Nuclei (DAPI, blue); scale bar, 1 mm.

(B) Relative mRNA levels of brown fat marker genes Ucp1 and Cidea. n=4 mice per group.

(C) Heat map of gene expression profiles comparing iWAT from young and aged mice, treated with vehicle or CL.

(D) Gene ontology (GO) and pathway analysis of a gene cluster downregulated by CL selectively in young vs. aged mice (red cluster, panel C). KG, KEGG Pathway; MF, Molecular Function; BP, Biological Process; CC, Cellular Component.

(E) Relative mRNA levels by RT-qPCR of fibrosis marker genes in iWAT. n=4 mice per group.

(F) Relative mRNA levels of fibrosis marker genes in stromal vascular cells (SVC) and mature adipocytes from young mice housed at thermoneutrality or exposed to 4°C for 1 week. n=3 mice per group.

(G) Relative mRNA levels of fibrosis marker genes in SVC from young and aged mice housed at thermoneutrality and treated with vehicle or CL for 4 days. n=3 mice per group.

(H) Picrosirius red staining of collagen fibers in iWAT from young and aged mice treated with vehicle (Ctl) or CL. Scale bar, 50 μM.

All data presented as mean ± s.e.m; *p<0.05, ** p<0.01, ***p<0.001 as analyzed by two-tailed Student’s t-test.